Difference between revisions of "Adipose Tissue Nuclear Isolation"
Davebridges (Talk | contribs) (copied initial protocol from Kang) |
(No difference)
|
Revision as of 20:41, 27 March 2017
From Kang et al
Epididymal adipose tissue was collected from chow- (n=15) and high-fat-fed (n=7) C57BL/6 mice. Pooled fat pads were minced and dounce homogenized with 10 strokes in hypotonic lysis buffer (10mM HEPES, pH 7.5, 10mM KCl, 1.5mM MgCl2 , 250mM sucrose, 0.5% NP40, and protease inhibitor cocktail). Lysates were filtered through a 100 µm cell strainer and spun at 1, 500g for 5 min. Lipid and cytoplasmic fractions were removed and the nuclear pellet was resuspended in lysis buffer
References
Kang S, Tsai LT, Zhou Y, Evertts A, Xu S, Griffin MJ, Issner R, Whitton HJ, Garcia BA, Epstein CB, Mikkelsen TS, Rosen ED. Identification of nuclear hormone receptor pathways causing insulin resistance by transcriptional and epigenomic analysis. Nat Cell Biol. 2015; 17: 44–56. doi: 10.1038/ncb3080.
